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UNIVERSITÉ DE SHERBROOKE Faculté De Génie Département De Génie Chimique Et Génie Biotechnologique

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UNIVERSITÉ DE SHERBROOKE Faculté De Génie Département De Génie Chimique Et Génie Biotechnologique UNIVERSITÉ DE SHERBROOKE Faculté de génie Département de génie chimique et génie biotechnologique UTILISATION DU LACTOSÉRUM DANS UN PROCÉDÉ DE CULTURE DE MICROALGUES MIXOTROPHES POUR LA PRODUCTION DE BIODIESEL Thèse de doctorat Spécialité : génie biotechnologique Jean-Michel Bergeron Girard Jury : Michèle Heitz (directrice) Jean-Sébastien Deschênes (co-directeur) Réjean Tremblay (co-directeur) Nathalie Faucheux (co-directrice) J. Peter Jones (rapporteur) Isabelle Marcotte (évaluatrice externe) Joël Sirois (évaluateur interne) Sherbrooke (Québec) Canada Septembre 2014 À ma mère, à mon père, à ma sœur et à Maud qui cultivent avec moi la légèreté du quotidien RÉSUMÉ L‘objectif général du présent travail est la conception et le développement d‘un procédé original de culture de microalgues pour la production à grande échelle d‘huiles végétales à faible coût pour le marché du biodiesel. Une procédure multicritères de sélection de souches a d‘abord été mise au point afin d‘identifier la souche la plus susceptible de répondre favorablement aux paramètres du procédé préalablement déterminés. Cette procédure permet d‘inclure un grand nombre de critères et de considérer l‘importance relative de chacun de ces critères dans le pointage final accordé aux souches présélectionnées. Elle peut aussi être transposée à d‘autres contextes techniques et géographiques pour la sélection de souches destinées à diverses applications commerciales. Par la suite, l‘évaluation de la croissance et de la productivité lipidique de la souche sélectionnée a été effectuée au laboratoire, en mode nutritionnel mixotrophe, en présence d‘un important coproduit de l‘industrie laitière : le perméat de lactosérum. Le profil lipidique de la souche privilégiée, ainsi que sa capacité à hydrolyser le lactose, mise au jour pour la première fois, ont permis de démontrer le potentiel du procédé. Une méthode de suivi de consommation et d‘hydrolyse du lactose contenu dans le perméat de lactosérum a été développée par l‘intermédiaire de la technologie « spectroscopie infrarouge à transformée de Fourier » couplée à une sonde externe avec « prisme à réflectance totale atténuée » (FT-IR/ATR) ainsi que la conception de modèles univariés et multivariés de type « régression des moindres carrés partiels (PLS) ». Ce type de modèle est plus précis que ceux déjà existants pour le suivi des sucres dissous par la méthode infrarouge car l'utilisation de régressions PLS permet de tenir compte du phénomène d‘additivité des spectres lorsque différents analytes pouvant absorber dans une même région du spectre sont présents dans la solution. De plus la méthode développée permet d‘envisager un suivi in situ de la consommation des sucres dans une culture de microorganismes à l‘échelle industrielle. La méthode des surfaces de réponse (RSM) a ensuite été utilisée afin d‘optimiser les conditions de pH et de densité de biomasse permettant de provoquer une accumulation de lipides à l‘intérieur de cultures de microalgues carencées en azote. Des modèles prédictifs ont été construits pour trois souches différentes et ce en modes nutritifs photoautotrophe et hétérotrophe. Ces travaux ont permis de démontrer l‘existence d‘une interaction entre ces deux facteurs. De plus, certaines différences spécifiques ont pu être mises en évidence en comparant les réponses entre les souches. À la lumière des résultats obtenus, certains paramètres d‘optimisation devront être considérés dans le cadre d‘un procédé de production de biomasse microalgale oléagineuse en deux phases; la première phase permettant d‘obtenir une productivité en biomasse maximale par une stratégie d‘augmentation du taux d‘hydrolyse du lactose; la seconde phase permettant une accumulation de lipides grâce à l‘imposition de conditions environnementales spécifiques. L‘augmentation des connaissances disponibles sur les microalgues ainsi que l‘amélioration et la standardisation des techniques de génie génétique nous permettront d‘aborder ces problématiques d‘un point de vue différent dans le futur. Mots-clés : Microalgue, lipides, biodiesel, lactosérum, mixotrophie i ii REMERCIEMENTS Je tiens à remercier mes directeurs et directrices de recherche pour ce projet : Jean-Sébastien Deschênes, Réjean Tremblay, Michèle Heitz et Nathalie Faucheux. Les autres personnes qui ont cordialement accepté de s‘impliquer : Jonathan Gagnon, Claude Belzile, Iften Redjah, Jean-Bruno Nadalini, Bertrand Genard, Pierre St-Onge, Alexandre Boudreau, Thomas Poirier- Audet, Mhammed Ben Hafsa, Alain Caron, Andréane Tétrault et Marie-Laine Roy. Le Fond Québécois de Recherche sur la Nature et les Technologies (FQRNT) pour le financement initial du projet, Ressources Aquatiques Québec (RAQ) et le Conseil de Recherche en Sciences Naturelles et en Génie (CRSNG) pour l‘aide financière fournie par l‘intermédiaire du programme de formation orientée vers la nouveauté, la collaboration et l'expérience en recherche (FONCER). Je tiens aussi à remercier tous les membres de ma famille, mes amis et Maud pour le soutien indéfectible tout au long de cette belle aventure. TABLE DES MATIÈRES RÉSUMÉ ...................................................................................................................................... i REMERCIEMENTS .................................................................................................................. ii TABLE DES MATIÈRES ......................................................................................................... iii LISTE DES FIGURES ............................................................................................................... vi LISTE DES TABLEAUX .......................................................................................................... ix LISTE DES ACRONYMES ET ABRÉVIATIONS .................................................................. xi CHAPITRE 1 Introduction ................................................................................................... 1 1.1 Mise en contexte et problématique ............................................................................... 1 1.2 Définition du projet de recherche ................................................................................. 3 1.3 Objectifs du projet de recherche ................................................................................... 4 1.4 Contributions originales ................................................................................................ 4 1.5 Plan du document .......................................................................................................... 6 CHAPITRE 2 État de l‘art .................................................................................................... 9 2.1 Sélection et amélioration de souches de microalgues ................................................... 9 2.1.1 Sélection sur plusieurs générations...................................................................... 10 2.1.2 Modification génétique ........................................................................................ 10 2.1.3 Biologie synthétique ............................................................................................ 11 2.2 Systèmes de production de microalgues ..................................................................... 11 2.2.1 PBRs tubulaires ................................................................................................... 13 2.2.2 PBRs colonnes ..................................................................................................... 14 2.2.3 PBRs panneaux plats ........................................................................................... 15 2.2.4 Autres modèles de PBRs ..................................................................................... 15 2.3 Modes de nutrition ...................................................................................................... 17 2.3.1 Photoautotrophie .................................................................................................. 17 2.3.2 Hétérotrophie ....................................................................................................... 17 2.3.3 Mixotrophie ......................................................................................................... 19 2.4 Bioraffinage ................................................................................................................ 20 2.4.1 Lipides ................................................................................................................. 20 2.4.2 Protéines .............................................................................................................. 26 iii iv 2.4.3 Hydrates de carbone ............................................................................................ 26 2.4.4 Applications commerciales de la biomasse microalgale entière ......................... 26 2.5 Biocarburants de troisième génération ....................................................................... 28 2.5.1 Bioéthanol et biométhane .................................................................................... 28 2.5.2 Biodiesel .............................................................................................................. 28 2.5.3 Propriétés du biodiesel ........................................................................................ 29 2.6 Valorisation du lactosérum ......................................................................................... 32 2.6.1 Utilisation du lactose
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